Fig 1.

Transformants of A. limacinum mh0186 and T. aureum ATCC 34304. (A) Schematic diagram of Ubi-Neo^r. Ubi-p, ubiquitin promoter from T. aureum ATCC 34304; Ubi-term, ubiquitin terminator from T. aureum ATCC 34304; Neo^r, neomycin resistance gene. (B) G418-resistant phenotypes of transformants generated by Ubi-Neo^r. Transformants were passaged 5 times in GY liquid medium without G418 and inoculated on PD agar plates containing G418 at the concentrations indicated. Incubation was carried out at 25°C for 7 days. The wild type was used as a control. (C) Fluorescence images of wild-type strains and transformants generated by Ubi-Neo^r showing lipid droplets. Lipid droplets were stained with Nile red as described in Materials and Methods. (D) Growth curves of wild-type strains and transformants generated by Ubi-Neo^r. Precultured cells were inoculated in 20 ml of GY liquid medium without G418 and incubated at 25°C with shaking at 150 rpm, and growth (optical density [OD] at 600 nm) was measured at the indicated times. Open circle, wild-type strain; closed circle, transfectant 1; closed triangle, transfectant 2; closed diamond, transfectant 3; asterisk, transfectant 4; plus mark, transfectant 5.